[Histonet] RE: Histonet Digest, Vol 21, Issue 2

From:

"Featherstone, Annette"

The Movat Pentachrome stain will give you your elastic, trichrome and alcian
blue. We use it all the time for our temporal biopsies.
Annette Featherstone HT/MLT
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
histonet-request@lists.utsouthwestern.edu
Sent: Tuesday, August 02, 2005 13:03
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 21, Issue 2
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than "Re: Contents of Histonet digest..."
Today's Topics:
1. Re: Problems of processing brain for paraffin embedding
(Gayle Callis)
2. New Lab Design (Breeden, Sara)
3. Results of "New Lab Ideas" Poll (Breeden, Sara)
4. Embedding human cartilage/joint tissue in plastic (Casey, Jane)
5. mouse anti-Her2 (Eva C Andersson)
6. Avoiding shrinkage Re: [Histonet] Embedding human
cartilage/joint tissue in plastic (Gayle Callis)
7. Histo tech position available (Cazares, Ruth)
8. Re: staining for iron in microglia (John Kiernan)
9. RE: New lab design (Malam Jacqueline)
10. Does anyone have a spare tissue processor that is getting in
the way? (Peter Bannister)
11. ANTI-MYOSIN AND ANTIRENIN (Jose Luis Palazon Fernandez)
12. retina neurons (Jose Luis Palazon Fernandez)
13. cassette labeller (Demarinis, Carolyn)
14. Elastic Trichrome - any good protocol out there not involving
microwave??? (GT Hebert)
15. RE: Elastic Trichrome - any good protocol out there no t
involving microwave??? (Monfils, Paul)
16. Re: Elastic Trichrome - any good protocol out there not
involving microwave??? (Bryan Hewlett)
17. RE: Elastic Trichrome - any good protocol out there no t
involving microwave??? (Bonner, Janet)
18. Whole Slide Imaging (Ben)
----------------------------------------------------------------------
Message: 1
Date: Mon, 01 Aug 2005 12:06:41 -0600
From: Gayle Callis
Subject: Re: [Histonet] Problems of processing brain for paraffin
embedding
To: azita parvaneh tafreshi ,
Histonet@lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050801120504.01b73828@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Are you processing whole brain, or coronal slices? Once you indicate, I
will be happy to send our protocols via private email attachement.
At 02:30 AM 7/30/2005, you wrote:
>Dear Members,
>I have problems with paraffin processing of mouse and hamster brains and
>spinal cords. I have perfused the animals with PFA 4% and I postfixed them
>in the same buffer for 7-8 days and then stored in Alc 70% (up to now).
>After sectioning, tissues wrinkle in a way that it is torn after
>flattenning (on a slide warmer), therefore the quality is not good at all.
>
>Can anyone give me hints to overcome the problem. A detailted protocol of
>paraffin processing would be great.
>
>Regard
>A.P. Tafreshi
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 2
Date: Mon, 1 Aug 2005 12:33:25 -0600
From: "Breeden, Sara"
Subject: [Histonet] New Lab Design
To:
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
Just a quick note to thank all of you that sent me your
ideas/comments/input on what to put into a new histo lab. These ideas
have been duly noted and will be part of My Grand Plan. There were some
meaty ideas and some real sparklers! Your help is invaluable -- and if
you've thought of anything else since last week, let me know. Thanks
again!
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
histonet-request@lists.utsouthwestern.edu
Sent: Monday, August 01, 2005 11:02 AM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 21, Issue 1
Send Histonet mailing list submissions to
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When replying, please edit your Subject line so it is more specific than
"Re: Contents of Histonet digest..."
Today's Topics:
1. Excessive background staining with Millers elastic stain
(Lachlan Smith)
2. staining for iron in microglia (Sharon Cooperman)
3. Re: Excessive background staining with Millers elastic stain
(Paul Bradbury)
4. RE: New Lab Design (Rogerson Kemlo (ELHT) Pathology)
5. Biotinylating Antibodies (Orr, Rebecca)
6. Re: EGFR (DDittus787@aol.com)
7. (no subject) (Patsy Ruegg)
8. RE: (no subject) (Patsy Ruegg)
----------------------------------------------------------------------
Message: 1
Date: Mon, 1 Aug 2005 09:33:17 +0930
From: "Lachlan Smith"
Subject: [Histonet] Excessive background staining with Millers elastic
stain
To:
Message-ID: <000001c5962c$6b43cca0$e46b140a@ITP36161>
Content-Type: text/plain; charset="iso-8859-1"
Dear Subscribers
I am staining cartilage using Millers elastic fibre stain, and while
elastic fibres are staining well, there is a lot of blue background
staining which makes thresholding for histoquantitation almost
impossible. My sections are 30 micron paraffin embedded. I am also
oxidising with 0.5% aqueous potassium permanaganate and bleaching with
2% oxalic acid.
Any suggestings for reducing this background staining would be greatly
appreciated.
Kind regards,
Lachlan Smith
Institute of Medical and Veterinary Science Adelaide, Australia
------------------------------
Message: 2
Date: Sun, 31 Jul 2005 21:39:40 -0400
From: Sharon Cooperman
Subject: [Histonet] staining for iron in microglia
To:
Message-ID:
Content-Type: text/plain; charset="us-ascii" ; format="flowed"
Dear Histonetters,
I have a semi-scientific question: macrophages in almost all tissues
can be stained for iron with Perl's stain to give a blue color - is this
also true for microglia? If not, is there any other way to see how much
iron is in microglia (DAB enhanced Perl's or some other method)?
Thanks,
Sharon
--
Sharon Cooperman
NIH, NICHD, CBMB 301.435-8417
Building 18T, room 101 301.402-0078 fax
Bethesda, MD 20892
------------------------------
Message: 3
Date: Sun, 31 Jul 2005 22:31:43 -0700
From: Paul Bradbury
Subject: Re: [Histonet] Excessive background staining with Millers
elastic stain
To: Lachlan Smith , HistoNet Server
Message-ID: <42EDB3BF.3030304@shaw.ca>
Content-Type: text/plain; format=flowed; charset=us-ascii
Hi Lachlan,
I believe the problem you are having is due to the presence of very
similar reactive groups in both cartilage and elastic fibres, Elastin
(the protein ) is heavily sulphated with an abundance of disulphide
groups (hence the yellowish appearance of elastic fibres when viewed
unstained or even macroscopically). When you oxidize the sections in
potassium permanganate, you are converting the disulphides to sulphates,
the sulphates react with Miller's elastin stain. Miller's stain is
essentially as modification of aldeyde fuchsin which iis a well known
method for demonstrating sulphated groups in mucopolysaccharides, etc.
The extracellular matrix of cartilage consists of chondroitin sulphate
(plus a few other assorted bits of connective tissue). These groups will
react quite strongly with Miller's stain solution.
So unfortunately, I think you may well be out of luck as far as
improving the specificity of the staining reactions. The same low
specificity reactions occur with most other elastin staining methods
when they are used on cartilage, the reactive groups present in both
locations stain with the dye in use. I cannot think of a method that
stains only elastin and not cartilage matrix.
Thinner sections may give you a better chance of distinguishing one from
the other. Try sections at several thinner settings and see what the
outcome is. Maybe there is a thickness at which you can see sufficient
enough detail in the elastic fibres without the cartilage obscuring it.
Paul Bradbury,
Kamloops, BC
Canada
Lachlan Smith wrote:
>Dear Subscribers
>
>I am staining cartilage using Millers elastic fibre stain, and while
>elastic fibres are staining well, there is a lot of blue background
>staining which makes thresholding for histoquantitation almost
>impossible. My sections are 30 micron paraffin embedded. I am also
>oxidising with 0.5% aqueous potassium permanaganate and bleaching with
>2% oxalic acid.
>
>Any suggestings for reducing this background staining would be greatly
>appreciated.
>
>Kind regards,
>
>Lachlan Smith
>Institute of Medical and Veterinary Science Adelaide, Australia
>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
------------------------------
Message: 4
Date: Mon, 1 Aug 2005 11:06:44 +0100
From: "Rogerson Kemlo (ELHT) Pathology"
Subject: RE: [Histonet] New Lab Design
To: "Breeden, Sara" ,
Message-ID:
<53A22BBB01D6184EBF7A4DC18A021E9E5AD4C0@elht-exch1.xelht.nhs.uk>
Content-Type: text/plain; charset="us-ascii"
AFOS (or something similar), formalin extraction for grossing. Includes
a downdraught table and formalin making up facilities. AFOS formalin
extraction cabinets for stored samples in formalin.
Be careful over height of benches; some are for sitting down carrying
out work, others for cutting sections using a microtome. Nothing worse
than a microtome that's too high or too low. You need solvent extraction
hoods for your processing machines, a suitable flammable store with
antiflash lighting etc., The best lighting to get it that stuff they
sell that goes in Offices; it's expensive but gives you a good colour
temperature with few shadows.
If you can see, stop people asphyxiating on formalin and alcohol, stop
the place blowing up and have benches the right height that people can
work, nowt else much to do. Oh yes, coffee machine, clean area and a
Mars Bar vending machine. Ergo, all done!
-----Original Message-----
From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu]
Sent: 29 July 2005 17:27
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Lab Design
A new histology lab (in a new building) is in the works (yipee!!). I
have some definite ideas about what I'll need in my new digs, but I need
more opinions! Within the next couple weeks, I will be asked by the
architects for my input.
This is a State veterinary lab where I do "surgicals", necropsies,
standard special stains, with a rapidly expanding IHC workload. My total
workload was roughly 10,000 blocks last year and I expect it to grow by
5% each year. I expect to have roughly 500 sq ft to work with in this
new space. I work alone for three DVM pathologists.
What would you consider absolutely essential in a new histology lab?
What would you do differently if you'd had a chance? What kind of: bench
space and type of surface, ventilation, lighting, hoods, haz mat
storage, supply storage, windows? I only get once chance at this and I
want to make it count...I have 8 more years until retirement and I'd
rather not kick myself for missing something obvious.
Please reply directly to me. Thank you in advance!! I rely on 'netters
for great information.
Sally Breeden, HT(ASCP)
New Mexico Department of Agriculture
Veterinary Diagnostic Services
POBox 4700
Albuquerque, NM 87196
(505)841-2576
(505)841-2518 FAX
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Mon, 1 Aug 2005 07:26:53 -0500
From: "Orr, Rebecca"
Subject: [Histonet] Biotinylating Antibodies
To:
Message-ID:
Content-Type: text/plain; charset="us-ascii"
Vernon,
I have been successful with a biotinylation kit from Biocare Medical.
It uses a different chemistry than the Vector kits.
I think the mopping reagent may be what makes the difference.
Their number is 800 799 9499 ask for Hari, he's the technical sales
manager.
Becky Orr, CLA HT (ASCP)
IHC Lead
Evanston Northwestern Healthcare
ph: 847-570-2771
------------------------------
Message: 6
Date: Mon, 1 Aug 2005 08:46:13 EDT
From: DDittus787@aol.com
Subject: Re: [Histonet] EGFR
To: JosefaNava@texashealth.org, histonet@pathology.swmed.edu
Message-ID: <9b.648c607f.301f7395@aol.com>
Content-Type: text/plain; charset="US-ASCII"
Josie:
Are you aware of the clone 31g7 and Allen Gowns ASCO 2005 poster. It
showed that 31g7 from Zymed was best in class and selected 10-15% more
patients.
dana
------------------------------
Message: 7
Date: Mon, 1 Aug 2005 10:21:23 -0600
From: "Patsy Ruegg"
Subject: [Histonet] (no subject)
To: "'Histonet'"
Message-ID: <200508011621.j71GLFpf017202@chip.viawest.net>
Content-Type: text/plain; charset="us-ascii"
Does anyone know where I could get a service manual for an old IEC
cryostat?
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg@ihctech.net
web site www.ihctech.net
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------------------------------
Message: 8
Date: Mon, 1 Aug 2005 10:23:37 -0600
From: "Patsy Ruegg"
Subject: RE: [Histonet] (no subject)
To: ,
Message-ID: <200508011623.j71GNTpf017949@chip.viawest.net>
Content-Type: text/plain; charset="us-ascii"
We used picro sirrus stain for collagen fibers and viewed the collagen
bundles using polarized light filter for histomorphometry.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg@ihctech.net
web site www.ihctech.net
This email is confidential and intended solely for the use of the
Person(s)
('the intended recipient') to whom it was addressed. Any views or
opinions
presented are solely those of the author. It may contain information
that is
privileged & confidential within the meaning of applicable law.
Accordingly
any dissemination, distribution, copying, or other use of this message,
or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited.
If
you are NOT the intended recipient please contact the sender and dispose
of
this e-mail as soon as possible.
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
lachlan.smith@imvs.sa.gov.au
Sent: Sunday, July 31, 2005 3:18 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] (no subject)
Dear Subscribers
I am staining cartilage using Millers elastic fibre stain, and while
elastic
fibres are staining well, there is a lot of blue background staining
which
makes thresholding for histoquantitation almost impossible. My sections
are
30 micron paraffin embedded. I am also oxidising with 0.5% aqueous
potassium
permanaganate and bleaching with 2% oxalic acid.
Any suggestings for reducing this background staining would be greatly
appreciated.
Kind regards,
Lachlan Smith
Institute of Medical and Veterinary Science Adelaide, Australia
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 21, Issue 1
***************************************
------------------------------
Message: 3
Date: Mon, 1 Aug 2005 13:17:21 -0600
From: "Breeden, Sara"
Subject: [Histonet] Results of "New Lab Ideas" Poll
To:
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
And with many thanks to all of you that had input, herewithin is a list
of the thoughts. And, might I add, this is just one reason I think this
medium (Histonet) is absolutely necessary!! Lookie what I found
out!!...
Large SINKS - and more than one; sink in HOOD for specials, IHC, etc.;
ergonomically correct SEATING and COUNTER HEIGHT for task; proper
VENTILATION and AIR DIRECTION (as in no airflow above microtome/water
bath, but proper ventilation for fume removal and room temperature
control); BRIGHT WHITE LIGHTING in task areas; VENT (ambient air
pressure?) or similar over embedding area/processor (for heat removal);
DEDICATED CIRCUITS for essential equipment and BACKUP POWER UNITS for
same; D.I. WATER; separate HAZMAT STORAGE and STORAGE for acids and
bases (separately); STORAGE for supplies/inventory (larger than you
think you'll ever need); EYEWASH/SHOWER; TEXTURED FLOOR; 30" deep
COUNTERS; rounded COUNTERTOP edges; COUNTERTOPS that don't stain or
corrode (stainless steel not recommended); no GLASS-INSERT DOORS
(visible clutter); overhead VENTS w/snorkels; and appropriate WORKFLOW
planning.
Of course, this prompted a few "needs" for my particular space, which I
will list just for informational purposes. But it may jog a thought for
someone in a similar position: HAZMAT storage out of work area or in an
area that's near the need (i.e., processor); trash cans out of walkway
(!); area for slide return (post-pathologist); locking cabinets and
drawers (for blades, knives, Secret Histology Stuff, etc.); and
under-cabinet lighting.
I'm sure the list will grow.
Sally Breeden, HT(ASCP)
New Mexico Department of Agriculture
Veterinary Diagnostic Services
POBox 4700 Albuquerque, NM 87196
(505)841-2576
(505)841-2518 FAX
------------------------------
Message: 4
Date: Mon, 1 Aug 2005 15:22:58 -0400
From: "Casey, Jane"
Subject: [Histonet] Embedding human cartilage/joint tissue in plastic
To:
Message-ID:
<471F15E271937A4DA89AC12211DDAD980266E9E3@MAIL2.AD.Brown.Edu>
Content-Type: text/plain; charset="iso-8859-1"
We are trying to use histology to validate our method of measuring cartilage
thickness on wrist bones using ?CT scans. Initially we tried fixing a
trapezoid fragment in formalin, de-mineralizing it, and embedding it in
paraffin before slicing it into 5 ?m sections. This method led to
non-uniform shrinkage of the tissue and clearly visible artifact. We would
like to try a method, perhaps embedment in plastic, which will yield a more
accurate measure of cartilage thickness. Does anyone have any suggestions?
Thank you for your suggestions.
Jane Casey
Department of Orthopaedics
Brown Medical School / Rhode Island Hospital
Providence, RI 02912
Jane_Casey@Brown.edu
------------------------------
Message: 5
Date: Mon, 01 Aug 2005 16:25:34 -0400
From: Eva C Andersson
Subject: [Histonet] mouse anti-Her2
To: histonet@lists.utsouthwestern.edu
Message-ID: <42EE853E.2010103@georgetown.edu>
Content-Type: text/plain; charset=us-ascii; format=flowed
Hello Everyone,
I just wanted to ask about a mouse anti-Her2 clone CB11 antibody that I
have bought from Zymed. It says the optimal dilutions are to be
determined by each lab but I just want to know if someone could tell me
which dilution they use. Also do you use an antigen retrieval method and
which detection method do you use?
Thank you
Eva
Georgetown University
------------------------------
Message: 6
Date: Mon, 01 Aug 2005 14:28:34 -0600
From: Gayle Callis
Subject: Avoiding shrinkage Re: [Histonet] Embedding human
cartilage/joint tissue in plastic
To: "Casey, Jane" ,
Histonet@lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050801141209.01b3ac10@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
A clever, simple use of low voltage xray machines i.e FAXITRON can help you
out.
We did specimen radiographs with a FAXITRON, and underexposed the sample to
enhance soft tissues i.e tumors and cartilage. These will be like
"contact" photographs, showing no shrinkage of the soft tissue
components. After fixation DO not decalcify, radiograph the sample then
decalcify (by the way, xray is the most sensitive decalcification endpoint
determination) and do another radiograph at the same initial exposure. You
can do thicker slabs even 5 mm thick but make sure you are consistent with
thickness of slab, a good bone saw will do this - Buehler Isomet or the
MarMed saw, a lovely device.
The bones can be NBF fixed but you avoid all dehydration/clearing and heat
of paraffin which will cause shrinkage. If you have to know exact edge of
cartilage, a radioopaque substance can be painted on surface of cartilage
to define that sharp edge.
There is a publication where a group tested the amount of shrinkage you
get with both paraffin and plastic processing, results were interesting in
that BOTH methods caused as much as 20% shrinkage, possibly more of the
hard tissue, so you will not avoid shrinkage with plastic embedding methods
involving alcohols or any other solvent to remove water.
Since cartilage has a high water content and by doing specimen
radiography, you can avoid alcohols, clearing agents and heat of paraffin
that cause shrinkage by water removal.
At 01:22 PM 8/1/2005, you wrote:
>
>
>We are trying to use histology to validate our method of measuring
>cartilage thickness on wrist bones using ?CT scans. Initially we tried
>fixing a trapezoid fragment in formalin, de-mineralizing it, and embedding
>it in paraffin before slicing it into 5 ?m sections. This method led to
>non-uniform shrinkage of the tissue and clearly visible artifact. We would
>like to try a method, perhaps embedment in plastic, which will yield a
>more accurate measure of cartilage thickness. Does anyone have any
>suggestions?
>
>Thank you for your suggestions.
>
>
>Jane Casey
>Department of Orthopaedics
>Brown Medical School / Rhode Island Hospital
>Providence, RI 02912
>Jane_Casey@Brown.edu
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 7
Date: Mon, 1 Aug 2005 16:09:48 -0500
From: "Cazares, Ruth"
Subject: [Histonet] Histo tech position available
To:
Message-ID:
<913FAC2B773C19488E26AE6572180FA5029A992C@exch01.schosp.org>
Content-Type: text/plain; charset="us-ascii"
Hi all,
We have a position open for a histo tech at Swedish Covenant Hospital in
Chicago. The position is alternate Saturdays (8 hour day) and fill in
for vacations if possible.
There is also opportunity to pick up extra hours during the week on
evenings but this is optional.
There will also be a full time position open in December when one of my
FT techs retires.
Interested techs (certified) can call or fax their resume;
Ruth Cazares
Department of Pathology
Phone: 773 878-8200 ext-5190
Fax: 773 271-1501 (to my attention)
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------------------------------
Message: 8
Date: Tue, 02 Aug 2005 00:16:09 -0400
From: John Kiernan
Subject: Re: [Histonet] staining for iron in microglia
To: Sharon Cooperman ,
histonet@lists.utsouthwestern.edu
Message-ID: <42EEF389.A65BF4AC@uwo.ca>
Content-Type: text/plain; charset=us-ascii
Sharon, Your question is 100% scientific,
not semi-scientific! You concisely provide an
introduction, expand it into a testable hypothesis,
and ask if the test has been done.
The answer is "Yes, but ..."
Iron is seen in phagocytic cells in various pathological
conditions of the brain. Its presence in the frontal
cortex (seen with the regular Perls method) is a
classical feature of advanced syphilis (general paralysis
of the insane). There are some cells in the normal rat's
hypothalamus that have a variety of histochemical
peculiarities including being Perls-positive, but these
cells are astrocytes (Young & McKenzie, 2004).
The DAB-enhanced Perls method shows non-haem iron in
microglia, oligodendroglia and neurons. Research in this
field, from several labs, was published in the late 1980s
and early 1990s, about 10 years after the publication of
the DAB amplification procedure by
Nguyen-Legros et al in 1980. A few references are given
at the end of this message. They are ones that I've seen
and made notes on; there will be many more, findable by
way of PubMed, Web of Science or Scopus. At NIH all
those resources and more will be available to you.
Another iron-related approach to microglia has been
immunostaining for ferritin (Kaneko et al 1989). There
are some lectins with affinity for microglia that can
provide impressive pictures when correctly applied.
For the old and bold there are the original silver
stains, developed by Del Rio Hortega and perfected by
Penfield and Cone in the 1920s.
Summary. Sensitive methods for iron do not provide
specific staining of resident, activated or immigrant
microglial cells. The classical Perls method can
detect microglia that have collected iron in sites of
injury or disease. Iron is also present in macroglia
and neurons, and is stainable in these cells if the
sensitivity of the Perls prussian blue method is
enhanced.
References.
Gerber MR, Connor JR (1989) Do oligodendrocytes mediate iron
regulation in the human brain? Ann. Neurol. 26: 95-98.
Kaneko Y, Kitamoto T, Tateishi J, Yamaguchi K (1989) Ferritin
immunohistochemistry as a marker for microglia. Acta Neuropathol.
(Berl.) 79: 129-136.
Morris CM, Candy JM, Oakley AE, Bloxham CA, Edwardson JA (1992)
Histochemical distribution of non-haem iron in the human brain.
Acta Anat. 144: 235-257.
Connor JR, Pavlick G, Karli D, Menzies SL, Palmer C (1995) A
histochemical study of iron-positive cells in the developing rat
brain. J. Comp. Neurol. 355: 111-123.
Palmer C, Menzies SL, Roberts RL, Pavlick G, Connor JR (1999)
Changes in iron histochemistry after hypoxic-ischemic brain
injury in the neonatal rat. J. Neurosci. Res. 56: 60-71.
Young JK, McKenzie JC (2004) GLUT2 immunoreactivity in
Gomori-positive astrocytes of the hypothalamus. J. Histochem.
Cytochem. 52: 1519-1524.
John Kiernan
Anatomy, UWO
London, Canada
---------------------------------------
Sharon Cooperman wrote:
>
> Dear Histonetters,
>
> I have a semi-scientific question: macrophages in almost all tissues
> can be stained for iron with Perl's stain to give a blue color - is
> this also true for microglia? If not, is there any other way to see
> how much iron is in microglia (DAB enhanced Perl's or some other
> method)?
>
> Thanks,
> Sharon
> --
> Sharon Cooperman
> NIH, NICHD, CBMB 301.435-8417
> Building 18T, room 101 301.402-0078 fax
> Bethesda, MD 20892
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 9
Date: Tue, 2 Aug 2005 08:41:07 +0100
From: Malam Jacqueline
Subject: [Histonet] RE: New lab design
To: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain
One thing you need to over-estimate on is block and slide storage, and if
you are not on a ground floor, they weigh a ton so you'll need some
additional support system.
Cheers
Jacqui Malam
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------------------------------
Message: 10
Date: Tue, 02 Aug 2005 12:01:29 +0100
From: "Peter Bannister"
Subject: [Histonet] Does anyone have a spare tissue processor that is
getting in the way?
To: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; format=flowed
Hello again Histonetters,
I realise that this is a slightly cheeky message, but I need to ask if any
of you (particularly those based in the UK) have an old carousel style
tissue processor taking up valuable storage space in your lab?
We are currently setting up a new research lab here in london that will have
a relatively low specimen throughput and a rather limited equipment budget.
We would therefore be eternally grateful if you would allow us to help you
dispose of your old kit!
We will of course collect and I will personally chip in towards your
christmas party.
Yes it's cheeky - but 'if you don't ask, you don't get!'
If you can help, please contact me directly. Many thanks for your time.
Peter Bannister
_________________________________________________________________
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------------------------------
Message: 11
Date: Tue, 2 Aug 2005 13:36:04 +0200 (CEST)
From: Jose Luis Palazon Fernandez
Subject: [Histonet] ANTI-MYOSIN AND ANTIRENIN
To: Histonet@lists.utsouthwestern.edu
Message-ID: <20050802113604.74D8A89C1F5@perceval.uca.es>
Content-Type: text/plain; charset="iso-8859-1"
Dear List-members
I am finishing my doctoral thesis and would like to demonstrate renin in an
aglomerular teleost kidney, smooth muscle in the iris, and muscle in the
bulbus arteriosus. I know that this can be done inmunohistologically but my
lab does not work in inmunohistochemistry so I do not have the antibodies. I
would like to know if there is someone of our list here in Spain (hospital,
University) that can do this kind of inmunos and the posibility of doing my
samples with them or maybe sending the samples and paying for the inmunos.
thanks in advance
José Luis
Universidad de Oriente-Isla Margarita-Venezuela
actualmente en: Instituto de Ciencias Marinas de Andalucia
Puerto Real, Cádiz, España.
email: jluis.palazon@icman.csic.es
------------------------------
Message: 12
Date: Tue, 2 Aug 2005 13:37:43 +0200 (CEST)
From: Jose Luis Palazon Fernandez
Subject: [Histonet] retina neurons
To: Histonet@lists.utsouthwestern.edu
Message-ID: <20050802113743.3F3E989C5A1@perceval.uca.es>
Content-Type: text/plain; charset="iso-8859-1"
Dear list fellows
is there any simple stain that I can use to differentiate between amacrine,
horizontal and bipolar cells in the retina. I have tried using H&E but as I
have not experience in this kind of tissue I dont know how to differentiate
between them, mainly between amacrine and bipolar cells.
thanks in advance
jose luis
Universidad de Oriente-Isla Margarita-Venezuela
actualmente en: Instituto de Ciencias Marinas de Andalucia
Puerto Real, Cádiz, España.
email: jluis.palazon@icman.csic.es
------------------------------
Message: 13
Date: Tue, 2 Aug 2005 07:43:35 -0400
From: "Demarinis, Carolyn"
Subject: [Histonet] cassette labeller
To: "HISTONET \(E-mail\)"
Message-ID:
Content-Type: text/plain; charset="iso-8859-1"
We are interested in obtaining a cassette labeller. Does anyone have any
recommendations? We are a Meditech hospital.
****************************************************************************
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------------------------------
Message: 14
Date: Tue, 2 Aug 2005 06:05:22 -0700 (PDT)
From: GT Hebert
Subject: [Histonet] Elastic Trichrome - any good protocol out there
not involving microwave???
To: histonet@lists.utsouthwestern.edu
Message-ID: <20050802130522.4981.qmail@web31710.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Hello all,
I am in need of a good protocol for a combination elastic and trichrome
stain that doesn't involve microwave. Thanks.
Gustave Hebert
Scientist II
Wyeth Research
Cambridge MA
__________________________________________________
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------------------------------
Message: 15
Date: Tue, 2 Aug 2005 09:52:22 -0400
From: "Monfils, Paul"
Subject: RE: [Histonet] Elastic Trichrome - any good protocol out
there no t involving microwave???
To: "'histonet@lists.utsouthwestern.edu'"
Message-ID:
<09C945920A6B654199F7A58A1D7D1FDE0171759D@lsexch.lsmaster.lifespan.org>
Content-Type: text/plain; charset="ISO-8859-1"
I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do
the Bouin's treatment required for the trichrome procedure first. Then I do
the complete Verhoeff stain, eliminatiing the final Van Gieson counterstain.
Then I apply the one-step Gomori Trichrome. It is quite simple and gives
excellent results.
Paul M.
> ----------
> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT
> Hebert
> Sent: Tuesday, August 2, 2005 6:05 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Elastic Trichrome - any good protocol out there
> not involving microwave???
>
> Hello all,
>
> I am in need of a good protocol for a combination elastic and trichrome
> stain that doesn't involve microwave. Thanks.
>
> Gustave Hebert
> Scientist II
> Wyeth Research
> Cambridge MA
>
> __________________________________________________
> Do You Yahoo!?
> Tired of spam? Yahoo! Mail has the best spam protection around
> http://mail.yahoo.com
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
------------------------------
Message: 16
Date: Tue, 2 Aug 2005 10:20:58 -0400
From: "Bryan Hewlett"
Subject: Re: [Histonet] Elastic Trichrome - any good protocol out
there not involving microwave???
To: "Monfils, Paul" ,
Message-ID: <01a301c5976d$67bdd250$6400a8c0@mainbox>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
Paul,
The Van Gieson counterstain IS a trichrome!
It also has the advantage of not requiring the Bouin pretreatment.
An excellent alternative to Verhoff, is the Miller elastic stain which also
incorporates the Van Gieson.
Bryan
----- Original Message -----
From: "Monfils, Paul"
To:
Sent: Tuesday, August 02, 2005 9:52 AM
Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there not
involving microwave???
>I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do
> the Bouin's treatment required for the trichrome procedure first. Then I
> do
> the complete Verhoeff stain, eliminatiing the final Van Gieson
> counterstain.
> Then I apply the one-step Gomori Trichrome. It is quite simple and gives
> excellent results.
>
> Paul M.
>
>> ----------
>> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT
>> Hebert
>> Sent: Tuesday, August 2, 2005 6:05 AM
>> To: histonet@lists.utsouthwestern.edu
>> Subject: [Histonet] Elastic Trichrome - any good protocol out there
>> not involving microwave???
>>
>> Hello all,
>>
>> I am in need of a good protocol for a combination elastic and trichrome
>> stain that doesn't involve microwave. Thanks.
>>
>> Gustave Hebert
>> Scientist II
>> Wyeth Research
>> Cambridge MA
>>
>> __________________________________________________
>> Do You Yahoo!?
>> Tired of spam? Yahoo! Mail has the best spam protection around
>> http://mail.yahoo.com
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 17
Date: Tue, 2 Aug 2005 10:31:15 -0400
From: "Bonner, Janet"
Subject: RE: [Histonet] Elastic Trichrome - any good protocol out
there no t involving microwave???
To: "'Bryan Hewlett '" ,
"'histonet-bounces@lists.utsouthwestern.edu '"
, "'Monfils, Paul '"
, "'histonet@lists.utsouthwestern.edu
'"
Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB434C@fh2k093.fhmis.net>
Content-Type: text/plain; charset=iso-8859-1
Generally, we find that the steps are so short anyway that we don't use the
microwave on the masson's trichrome-except for the Bouin's Solution and even
then we try not to because the solution becomes compromised.
---Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
To: Monfils, Paul; histonet@lists.utsouthwestern.edu
Sent: 8/2/2005 10:20 AM
Subject: Re: [Histonet] Elastic Trichrome - any good protocol out there not
involving microwave???
Paul,
The Van Gieson counterstain IS a trichrome!
It also has the advantage of not requiring the Bouin pretreatment.
An excellent alternative to Verhoff, is the Miller elastic stain which
also
incorporates the Van Gieson.
Bryan
----- Original Message -----
From: "Monfils, Paul"
To:
Sent: Tuesday, August 02, 2005 9:52 AM
Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there
not
involving microwave???
>I often combine the Verhoeff Elastin Stain with the Gomori Trichrome.
I do
> the Bouin's treatment required for the trichrome procedure first.
Then I
> do
> the complete Verhoeff stain, eliminatiing the final Van Gieson
> counterstain.
> Then I apply the one-step Gomori Trichrome. It is quite simple and
gives
> excellent results.
>
> Paul M.
>
>> ----------
>> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT
>> Hebert
>> Sent: Tuesday, August 2, 2005 6:05 AM
>> To: histonet@lists.utsouthwestern.edu
>> Subject: [Histonet] Elastic Trichrome - any good protocol out there
>> not involving microwave???
>>
>> Hello all,
>>
>> I am in need of a good protocol for a combination elastic and
trichrome
>> stain that doesn't involve microwave. Thanks.
>>
>> Gustave Hebert
>> Scientist II
>> Wyeth Research
>> Cambridge MA
>>
>> __________________________________________________
>> Do You Yahoo!?
>> Tired of spam? Yahoo! Mail has the best spam protection around
>> http://mail.yahoo.com
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
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------------------------------
Message: 18
Date: Tue, 2 Aug 2005 09:03:42 -0700
From: "Ben"
Subject: [Histonet] Whole Slide Imaging
To:
Message-ID: <200508021603.j72G3gUj002726@pop19.ucdavis.edu>
Content-Type: text/plain; charset="us-ascii"
Hello All,
I'm interested in whole slide imaging for some research work. I was hoping
that anyone who has worked with it could give me their opinion on the
equipment they use, the important features (by the way I'm looking to count
nuclei), or suggest a good place to get this done for a decent price. If
this message is against the rules of histonet, I apologize, but if not I
appreciate your help.
Ben Renquist
UC Davis
------------------------------
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End of Histonet Digest, Vol 21, Issue 2
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